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Title: Using peptide libraries to search for the substrates of ABH1 and Ofd2
Author: Balmer, J. N.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 2010
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ABH1 and Ofd2 are human and Schizosaccharomyces pombe members of the 2-oxyglutarate-iron(II)-dependent dioxygenase family respectively. Little is known about these proteins, and no substrates for them have been identified. Using positionally addressable synthesis of peptides on a cellulose membrane (SPOT synthesis) random peptide libraries were generated. These were screened with ABH1 and Ofd2 using a 2-oxoglutarate turnover assay, to generate preferred sequence motifs. The motifs were searched against protein databases to create a list of potential substrates. The potential substrates were tested both for binding and activity. Several highly active peptides were systematically mutated in an attempt to further define the preferred motifs. These sequence preferences were used to conduct another search of the protein databases. Ultimately none of the sequences studied could be confirmed as substrates, however, refinement of the technique used could produce a viable method of identifying unknown substrates.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available